扁玉螺体表和消化系统甲硫氨酸脑啡肽免疫组织化学定位

采用免疫组织化学strept avidin-biotin complex(S-ABC)法对甲硫氨酸脑啡肽(methionine-enkephalin,M-ENK)在扁玉螺(Neverita didyma)体表、消化系统各器官中的分布进行了研究.结果表明,扁玉螺的足上皮细胞、外套膜的内外上皮细胞以及食道、胃、肠道的黏膜上皮细胞均呈M-ENK阳性反应,且消化道中的阳性反应多集中于上皮细胞游离端;在食道腺的腺上皮中也有少量阳性细胞分布;肝是M-ENK阳性细胞分布较多的器官,主要分布在肝小叶中腺细胞边缘游离端.M-ENK在扁玉螺体表和消化系统各器官均有分布,且分布密度有所不同,可能与各部位的功能有关.     

华贵栉孔扇贝♀×栉孔扇贝♂远缘杂交的受精细胞学观察

用HOECHST33258对华贵栉孔扇贝(Chlamys nobilis)♀×栉孔扇贝(Chlamys farreri)♂的受精卵进行染色,在荧光显微镜下观察其受精细胞学过程。观察表明:栉孔扇贝的精子能够使华贵栉孔扇贝卵子受精。精子入卵后呈一圆形亮点,体积稍有膨大成球形;授精后成熟卵母细胞释放出第一极体和第二极体后,精核解凝、稀疏、泡状化,形成雄性原核(male pronucleus);雌性原核(female pronucleus)在完成两次成熟分裂之后,染色质去浓缩,扩散膨大;雌雄原核相互靠拢,当雌雄原核膨胀到最大程度时,发生融合,形成合子。受精卵能够正常发育,完成第一次卵裂,卵裂时细胞核分离未观察到异常现象。与华贵栉孔扇贝和栉孔扇贝种内交配对照组相比,异源受精卵的细胞学过程明显滞后,同时其发生过程具明显的不同步现象。实验中还观察到少数的雌核发育现象。     

柄袋沙蠋体壁和消化系统甲硫氨酸脑啡肽免疫组织化学定位

采用免疫组织化学SABC法,对甲硫氨酸脑啡肽(M-ENK)在柄袋沙蠋体壁及其附属物鳃、疣足和消化系统各器官中的分布进行了研究.结果 表明,柄袋沙蠋体壁、鳃、疣足以及各消化器官的上皮细胞中均有不同程度的M-ENK阳性反应,且体壁、消化道的食管、肠、直肠中阳性反应多集中在上皮细胞的游离端;体壁肌肉中环肌层M-ENK阳性反应比纵肌层强;食道侧囊上皮细胞和结缔组织中也有M-ENK分布.M-ENK在柄袋沙蠋的体壁、鳃、疣足及消化系统各器官中均有分布,且分布密度不同,可能与各部位的功能有关.     

四十里湾栉孔扇贝清滤率、摄食率和吸收效率的现场研究

运用生物沉积法在四十里湾不同海区对栉孔扇贝的生理生态学特征进行了研究。 1龄栉孔扇贝 (4 1 .1± 4.1 mm,软体干重 0 .48± 0 .1 0 g/ ind)清滤率变化范围为 0 .72～ 2 .5 4(平均 1 .2 7) L/ (ind· h)或 1 .65～ 5 .97(平均 2 .61 ) L/ (g· h)。清滤率受 TPM的变化影响不大 ,而摄食率却随 TPM的升高而升高。 2龄扇贝 (软体干重 1 .91± 0 .3 2 g/ ind)清滤率为2 .0 9～ 3 .99(平均 3 .1 0 ) L/ (ind· h)。栉孔扇贝吸收速率与 POM呈正相关关系 ,而与饵料质量 (POM/ TPM)无明显的相关性。1龄扇贝和 2龄扇贝吸收效率没有显著差别 ;扇贝对 POM的吸收效率与 TPM (或 POM)关系不大 ,却与饵料质量呈明显的正相关关系 ;扇贝对 POC、PON和 PP的吸收效率平均分别为 68.9%、64.0 %和 63 .6%。在沿岸养殖海域 ,栉孔扇贝通过大量的滤水摄食以及较高的吸收效率对生态系统的能量流动和物质循环产生影响。     

栉孔扇贝感染急性病毒性坏死症病毒的组织病理学与免疫荧光检测

应用组织学和单克隆抗体免疫荧光技术(Immunofluorescence assay,IFA)对夏季养殖中后期大规模死亡(“急性病毒性坏死症”Acute virus necrobiotic disease,AVND)高峰期的患病栉孔扇贝(Chlamys,farrerl)进行了检测。组织学检测结果显示,患病扇贝的大多数器官(外套膜、鳃、胃、肾等)的上皮组织细胞可见显著的细胞肿胀、嗜碱性增强、排列紊乱、部分脱落以至完全坏死脱落等显著的组织病理学变化。作为对照,利用针对AVND病毒的特异性单克隆抗体所建立的免疫荧光原位检测技术对患病扇贝进行检测发现,上皮组织的病理变化与病毒感染之间具有一致的对应关系。这一结果表明,AVND病毒的感染可以对栉孔扇贝造成严重的病理性破坏。此为解释该病毒导致栉孔扇贝大规模死亡的机制提供了直接的组织病理学依据。     

栉孔扇贝耗氧率和排氨率的研究

1999年 4～ 6月 ,采用室内实验生态学方法对栉孔扇贝的耗氧率和排氨率进行了研究 .结果表明 ,在适宜的温度范围内 ,栉孔扇贝的耗氧率和排氨率均与温度成正比 ,而与体重呈负相关关系 .在实验室温度 (8～ 2 8℃ )条件下 ,栉孔扇贝的耗氧率为 0 .48～ 9.0 9mg·g-1·h-1,排氨率为 0 .0 5～ 1 0 1mg·g-1·h-1.其中耗氧率在 2 3℃时达到最高值 ,2 8℃时开始下降 ,而排氨率则呈持续升高趋势 .栉孔扇贝的日常代谢明显高于标准代谢 ,耗氧率和排氨率平均值分别提高约 35 .8%和 75 .9% .     

栉孔扇贝生理生态学特征的实验研究

基于近年发展起来的生物沉积法,在烟台四十里湾养殖海区采用室内大型流水系统对栉孔扇贝的滤水率、吸收率和生态效率等生理生态学参数进行了测定,并探讨了它们与养殖密度与食物可获得性的关系．栉孔扇贝滤水率(平均3．65L·ind^-1·h^-1)与扇贝放养密度和饵料浓度无显著关系,而栉孔扇贝摄食率随放养密度的升高而降低,与POM呈正相关．扇贝对食物的吸收效率较高(平均75．9％),这与低饵料浓度行关．扇贝氨基酸泄漏所损失的能量高于排氨的能量损失．扇贝的生长余力(SFG)、总生长效率(尺：)和净生长效率(尺z)均与饵料浓度呈正相关．扇贝对氮的总生态效率(平均9．9％)高于对碳(平均5．9％)和磷(平均4．1％)．在氟的预算中,如果仅考虑NH4^ —N的排泄,而忽视其它形态氟的排泄,将会产生较大偏差(平均约20％)．贝壳不管在能量预算还是在元素预算中部不应该被忽视．在沿岸养殖海域,栉孔扇贝通过大量的滤水摄食、生长、排泄、生物沉积等对生态系统的能量流动和物质循环产生影响．     

硫化物对栉孔扇贝线粒体的损伤研究

硫化物是含有-2价硫离子化合物的统称,会抑制复合体IV活性,是水产养殖中常见的有毒物质。以水产养殖动物栉孔扇贝为研究对象,评估硫化物对线粒体的损伤。研究结果显示硫化物能显著降低栉孔扇贝各组织线粒体活力和SOD酶活,增加线粒体的肿胀度和MDA含量,影响线粒体的功能,造成线粒体的损伤,进而对栉孔扇贝产生毒害作用。     

不同水深养殖栉孔扇贝的形态测量学研究和比较

为了探究远岸深水海区和近岸浅水海区养殖的栉孔扇贝(Chlamys farreri)形态上的差异, 运用传统形态测量学和几何形态测量学的方法, 对来自近岸浅水海区(15 m水深)和远岸深水海区(30 m水深)底栖养殖的2龄栉孔扇贝进行形态测量学研究和比较分析。结果表明: 深水养殖的栉孔扇贝的壳高(SH)、壳长(SL)和壳宽(SW)都极显著大于浅水养殖的栉孔扇贝(P<0.01); 通过界标点(Landmarks)和半界标点(Semi-landmarks)的方法, 及广义普鲁克分析(GPA)、主成分分析(PCA)和典型变量分析(CVA), 结果表明深水养殖的栉孔扇贝的整体壳形尺寸大于浅水养殖的栉孔扇贝, 除去尺寸大小因素后两者的壳耳和壳体扇形边缘存在显著差异。文章为栉孔扇贝深水养殖的选育和增养殖工作提供基于形态学研究的理论依据, 同时比较分析了传统形态测量学和几何形态测量学的原理和分析方法, 为研究生物体形态差异时选择合适的研究方法提供新参考。     

用流式细胞仪测定扇贝血细胞吞噬活性

采用流式细胞仪技术对栉孔扇贝(Chlamysfarreri)和海湾扇贝(Argopectenirradians)血细胞吞噬活性进行了测定。通过对缓冲体系的筛选和比较,建立了扇贝血细胞吞噬活性的流式细胞仪检测方法,并应用该技术对两种扇贝血细胞的吞噬活性进行了比较研究。结果发现在TBS(0.05mol.L-1Tris-HCl,pH7.4;2%glucose;2%NaCl;20mmol.L-1EDTA)和PBS体系中,扇贝血细胞的吞噬活性几乎完全受到抑制,在CMPBS(0.1mol.L-1PBS,pH7.4;2%NaCl;2%glucose;1.0mmol.L-1CaCl2;0.5mmol.L-1MgCl2)体系中细胞吞噬活性略有升高,而在海水缓冲液中细胞吞噬活性最高,其中海湾扇贝和栉孔扇贝血细胞的吞噬率分别达到了26.73%和19.89%,且海湾扇贝血细胞的吞噬率显著高于栉孔扇贝(P<0.05)。研究结果为进一步探讨扇贝血细胞吞噬功能提供了方法依据。     

Immunohistological Detection of Leucine-Enkephalin in the Digestive System of the Scallop <Emphasis Type="Italic">Chlamys farreri</Emphasis>

The study was designed to determine whether leucine-enkephalin (L-ENK) was present in the digestive system of the scallop Chlamys farreri. The results indicate that L-ENK was present in the epithelium and connective tissue of mouth labia, labial palps, intestine, rectum, and stomach of the scallop Chlamys farreri. Moreover, it was also found that isolated cells showing L-ENK immunoreactivity were detected between the epithelial cells and the basal lamina in the principal hepatopancreatic ducts, and a few immunoreactive cells and fibers were observed between the hepatopancreatic tubules. Our report constitutes the first characterization of L-ENK in the digestive system of the scallop Chlamys farreri, and demonstrates its origin in simpler animals.     

Immunohistochemical Identification of Met-Enkephalin in Digestive System and Its Effect on Digestive Enzyme Activities of the Scallop <Emphasis Type="Italic">Chlamys farreri</Emphasis>

The study was designed to determine whether methionine-enkephalin (M-ENK) was present in the digestive system of the scallop Chlamys farreri and investigate the effects of M-ENK on the activity of amylase, protease and lipase in the digestive system of C. farreri. The results indicated that M-ENK-like material was present in the epithelium and connective tissue of labial palps, mouth labia, stomach, intestine, rectum, and hepatopancreas of the scallop C. farreri. Moreover, it was also found that many isolated small cells showing M-ENK-like immunoreactivity were scattered in the epithelial layer of intestine, and many isolated big epithelial cells showing M-ENK-like immunoreactivity were scattered in tubules of hepatopancreas of the scallop. The activity levels of amylase and lipase in crystalline style, hepatopancreas and intestine were enhanced at 1 h after injection of exogenous M-ENK into adductor muscle of the scallops, whereas protease activity levels were significantly suppressed. Our report constitutes the first characterization of M-ENK in the digestive system of scallop C. farreri and investigates the effects of M-ENK on the activities of digestive enzymes of mollusk for the first time. The results suggest an involvement of M-ENK in the functional regulation of the digestive system of C. farreri.     

刀鲚幼鱼消化系统的组织形态学结构

采用光镜和扫描电镜观察长江刀鲚(Coilia nasus)幼鱼消化系统组织形态学结构。结果显示,刀鲚体长,口裂大,含有犬齿状的颌齿和尖锥状的腭齿,具有5对鳃弓,鳃耙长度明显大于鳃丝且表面附着不规则绒毛状细齿;胃呈"Y"型,胃与肠连接处具有16～21个指状环形幽门盲囊;肠为直肠,较短,比肠长为0.241±0.080;肝分为两叶,胰为独立的器官。刀鲚口咽腔为复层鳞状上皮,含有腺体、大量椭圆形黏液细胞、少量杯状细胞及味蕾;胃黏膜都为典型的单层柱状上皮,含有较多由上皮凹陷形成的胃小凹和胃腺;幽门盲囊具有20～25个丰富的褶皱,占满大部分幽门盲囊腔,黏膜层具有微绒毛;中肠黏膜上皮最发达,形成的褶皱细长且连接成网状,单层柱状上皮与复层扁平上皮交替分布。观察结果表明,刀鲚消化系统具有典型肉食性鱼类特征。     

Fasciola gigantica: Histology of the digestive tract and the expression of cathepsin L

The digestive tract of Fasciola gigantica is composed of the oral sucker, buccal tube, pharynx, esophagus, and caecum. The tegumental-type epithelium lines the first four parts of the digestive tract while the caecal-type epithelium lines the remaining parts from the caecal bifurcation. The caecal-epithelial cells are classified into 3 types according to their staining properties and ultrastructural characteristics, as related to the amount of food contents in the caecal lumen. All caecal-type epithelial cells synthesize and secrete cathepsin L, a major group of enzymes in the digestive tract, as detected by in situ hybridization and immunolocalization. Moreover, the secreted cathepsin L is also adsorbed on the outer surface of the tegument and the glycocalyx coating of the surface of the tegument, whereas the tegumental cells and tegumental syncytium covering the parasite’s body and lining the proximal part of the digestive tract exhibit no in situ hybridization signal and immunostaining for cathepsin L.     

龟足消化系统形态和组织学特点

应用光学显微镜观察龟足（Capitulum mitella）消化系统的形态和组织结构。龟足的消化系统包括消化腺和消化道。消化腺一对,呈长囊状,含有分泌细胞（B细胞）、吸收细胞（R细胞）、储存细胞（F细胞）和胚细胞（E细胞）4种类型细胞。消化道呈U型,由口、食道、胃、肠、直肠和肛门组成,各部分的结构由内到外可分为黏膜层、黏膜下层、肌层和外膜4层。口器为咀嚼型,包括一片上唇、一对触须、一对大颚以及两对小颚。食道细短,具几丁质层但无基膜,管壁向腔内突起形成明显的纵褶突;食道前段的环肌特别发达,同时独有放射肌。胃略呈球袋状,肠较长;胃和肠的组织结构相似,没有几丁质层,上皮细胞都有发达的微绒毛。直肠细长,外膜分布有16组纵肌;直肠前段的组织结构与胃、肠相似,而直肠后段有几丁质层覆盖,黏膜层、黏膜下层、肌层和外膜渐退化,16组纵肌渐发达。肛门16组更加发达的纵肌挤入上皮细胞下方,在外膜外另出现一层明显的环肌。龟足消化道各部分的组织结构差异明显,反映了它们功能的差异。     

Application of monoclonal antibody against granulocytes of scallop Chlamys farreri on granulocytes occurrence at different developmental stages and antigenic cross-reactivity of granulocytes in five other bivalve species

A monoclonal antibody (MAb) 6H7 raised specifically against granulocytes of scallop (Chlamys farreri) was employed to observe granulocyte occurrence successively in blastulae, gastrulae, trochophore larvae, D-shape larvae, umbo-veliger larvae and creeping larvae of C. farreri by immunohistochemistry assay contrasted with H&E stain using semi-thin sections. Moreover, the reactivity of the MAb with granulocytes of C. farreri, Bay scallop Argopecten irradians, Japanese scallop Patinopecten yessoensis, Blue mussel Mytilus edulis, Pacific oyster Crassostrea gigas and Manila clam Ruditapes philippinarum, was detected by immunofluorescence assay (IFA) with differential interference contrast and fluorescent microscopy and flow cytometric immunofluorescence assay (FCIFA). The results showed that positive signals were first observed at D-shape larval stage, about 28 h post fertilization, after that, umbo-veliger larvae exhibited the positive cells with a diameter of 3–5 μm distributed in velum, digestive gland and esophagus. Then in creeping larvae, the number of positive cells increased with average diameter of 5–7 μm, and widely distributed in foot, digestive gland, gills and adductor muscles. No positive signal was found in blastulae, gastrulae and trochophore larvae. The results of IFA and FCIFA showed MAb 6H7 reacted to granulocytes of C. farreri, A. irradians, P. yessoensis and C. gigas, and the positive percentage reactivity were 53 ± 2.5%, 15 ± 2.5%, 12 ± 2.1% and 19 ± 2.1%, respectively, however, no cross-reaction was detected in hemocytes of R. philippinarum and M. edulis.     

野生与养殖黄鳍鲷消化道形态组织结构

采用常规石蜡组织切片的方法对野生和养殖黄鳍鲷(Sparus latus)消化道的形态组织结构进行了比较观察。结果表明,野生和养殖黄鳍鲷的消化道存在一定差异。(1)形态学研究表明,食道粗而短,胃呈V形,分为贲门部、胃体部和幽门部,胃与肠的连接处有4条幽门盲囊,肠道在体腔内迂回两个回折。野生黄鳍鲷牙齿更为坚硬锋利,体腔中脂肪较少,消化道更为粗短。野生和养殖黄鳍鲷的肠道系数分别为0.71±0.03和0.94±0.12。(2)组织学研究表明,食道黏膜上皮由扁平细胞层和杯状细胞层组成,杯状细胞发达。胃黏膜由单层柱状上皮组成,无杯状细胞,贲门部和胃体部胃腺发达。幽门盲囊组织学特征与肠相似,上皮为柱状上皮,其中的杯状细胞少于肠。肠中,前肠杯状细胞最多,中肠次之,后肠最少。直肠杯状细胞多于肠。野生与养殖黄鳍鲷组织学的区别在于,消化道相同部位养殖鱼的杯状细胞多于野生鱼,野生鱼的肌层厚度大于养殖鱼。黄鳍鲷消化道的形态组织结构与其生活环境和食物是相关的。     

NADPH-diaphorase activity in the digestive system of gastropod molluscs <Emphasis Type="Italic">Achatina fulica</Emphasis> and <Emphasis Type="Italic">Littorina littorea</Emphasis>

Distribution and morphological peculiarities of nitroxidergic elements throughout the entire length of digestive tract was studied for the first time in gastropod molluscs Littorina littorea (Prosobranchia) and Achatina fulica (Pulmonata) using histochemical detection of NADPH-diaphorase (NADPHd). NO-ergic cells and fibers were revealed in all parts of the mollusc digestive system beginning from esophagus. Intensive NADPHd activity is found in a great number of intraepithelial cells of the open type and their processes in the intraand subepithelial nerve plexuses, subepithelial neurons, granular connective tissue cells, and multiple nervous fibers distributed among muscular elements of digestive tract as well as those in nerves innervating the tract. NADPHd was also revealed in receptor cells in the oral area and in the A. fulica CNS ganglia innervating the digestive tract. A. fulica has a more complicated organization of A. fulica nitroxidergic system of the digestive tract. A system of glomerular structures formed by thin NADPHd-positive neural fibers coming from epithelium is found directly beneath the epithelium in esophagus, stomach, and midgut of the mollusc. More superficially under the main groups of muscular elements there are revealed small clusters of NADPHd-positive neurons that can be classified as primitive, non-structured microganglia. The distribution pattern and a possible functional role of nitroxidergic elements in digestive tract of molluscs as compared with other invertebrate and vertebrate animals are discussed.     

GABA-immunoreactive structures in rat kidney.

We examined the distribution of gamma-aminobutyric acid-like immunoreactivity (GABA-LI) in the rat kidney by light and electron microscopy. In vibratome sections, GABA-LI was present in both the renal medulla and cortex. The inner stripe of the outer medulla was most heavily and almost homogeneously labeled, whereas GABA-LI in the cortex was mainly confined only to some tubules. GABA-positive structures involved the epithelial cells of the thin and the thick ascending limbs of the loop of Henle, the connecting tubules, and the collecting ducts. In GABA-positive connecting tubules and collecting ducts the immunoreactivity was present in the cytoplasm of about half of the epithelial cells. As revealed by electron microscopy, the labeled cells in the collecting tubules were the light (principal) cells. No GABA-LI occurred in neuronal structures. These findings are consistent with the presence of a non-neuronal GABA system in the rat kidney. Furthermore, the specific distribution of GABA in the tubular epithelium suggests a functional significance of this amino acid in tubular transport processes.     

黑麂肝、小肠和大肠的组织学结构及Ghrelin的分布

研究了成年雌性黑麂(Muntiacus crinifrons)的肝、小肠和大肠的组织学结构及Ghrelin的分布。采用H.E染色法观察组织学结构,免疫组化PV-9000两步法并结合DAB显色技术确定Ghrelin的分布。结果表明,黑麂的肝组织分为被膜、肝小叶、肝中央静脉、门管区等结构。被膜为浆膜结构,肝小叶不明显。肝细胞以中央静脉为中心,呈放射状排列。肝血窦的形状不规则。肠黏膜上皮为单层柱状上皮,小肠、盲肠和结肠的黏膜肌层很薄,管壁皱襞与肠绒毛等形态在消化道各部也存在差异。免疫组化结果显示肝细胞中有Ghrelin阳性细胞的表达;在肠道,免疫阳性细胞在十二指肠、空肠、回肠、盲肠和直肠的黏膜、黏膜下层和肌层均有分布,尤其在肠绒毛上皮和黏膜下层分布较多。黑麂肝、小肠和大肠结构与哺乳动物基本相似,但无十二指肠腺;Ghrelin阳性细胞在肝、小肠和大肠均有分布,这表明Ghrelin可能对消化有一定的调节作用。